Mouse anti-vinculin (V9131, dilution ratio: 1/20,000 (WB), 1/500 (IF)), rabbit anti-CAP (SORBS1) (HPA027559, 1/2,000 (WB), 1/100 (IF)) and anti-β tubulin (T4026, 1/2,000) antibodies were purchased from Sigma (Saint Louis, MO). Mouse anti-β-actin antibody (ab6276, 1/10,000) was purchased from Abcam (Cambridge, UK). Mouse anti-YAP (sc-101199, 1/100 (IF), 1/1000 (WB)), and rabbit anti-ERK2 (sc-154, 1/10,000) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit anti-aP2 (#3544, 1/2,000) antibodies was purchased from Cell Signaling Technology (Boston, MA). Rabbit anti-vinexin and anti-ArgBP2 polyclonal antibodies were described previously18 (link),22 (link). Alexa Fluor 568 phalloidin and 633 phalloidin was purchased from Thermo Fisher Scientific (Rockford, IL). Type I collagen was purchased from Nitta Gelatin (Osaka, Japan). Insulin and 3-Isobutyl-1-methylxanthine (IBMX) were purchased from Sigma.
Anti β tubulin t4026
Manufactured by Merck Group
Sourced in United States
Anti-β tubulin (T4026) is a laboratory reagent used for the detection and localization of β-tubulin in cells and tissues. It functions as a specific antibody that binds to β-tubulin, a key component of the cytoskeleton. This product can be used in various cell biology and biochemical applications, such as Western blotting, immunocytochemistry, and immunohistochemistry.
Automatically generated - may contain errors
Lab products found in correlation
3 isobutyl 1 methylxanthine ibmx, by Merck Group (1 mentions)
Mouse anti yap sc 101199, by Santa Cruz Biotechnology (1 mentions)
Mouse anti vinculin v9131, by Merck Group (1 mentions)
Mouse anti β actin antibody ab6276, by Abcam (1 mentions)
Alexa fluor 568 phalloidin, by Thermo Fisher Scientific (1 mentions)
Insulin, by Merck Group (1 mentions)
Type 1 collagen, by Nitta Gelatin (1 mentions)
Ab32034, by Abcam (1 mentions)
Plx 4720, by MedChemExpress (1 mentions)
Anti vinculin v4505, by Merck Group (1 mentions)
Otx 008, by Axon Medchem (1 mentions)
Branson sonifier 450, by Emerson (1 mentions)
Myhc mf20, by Developmental Studies Hybridoma Bank (1 mentions)
Anti myogenin sc 12732, by Santa Cruz Biotechnology (1 mentions)
Amersham imagequant 800, by GE Healthcare (1 mentions)
Pierce ecl western blotting substrate, by Thermo Fisher Scientific (1 mentions)
Anti β actin, by Merck Group (1 mentions)
Ecl western blotting detection reagent, by GE Healthcare (1 mentions)
Anti acetylated α tubulin t7451, by Merck Group (1 mentions)
Las4000, by GE Healthcare (1 mentions)
7 protocols using anti β tubulin t4026
1
Immunoblotting and Immunofluorescence Protocols
2
Antibody Detection of EGFR and p27/Kip1
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EGFR was detected with an antibody purchased from Enzo Life Sciences, Farmingdale, NY, USA (cat. ALX-804-064-C100). The p27/Kip1 antibody (ab32034) was purchased from Abcam, Cambridge, UK. Other antibodies applied in this study were anti-vinculin (V4505, Sigma) and anti-βtubulin (T4026, Sigma). Secondary antibodies were purchased from Promega (Madison, WI, USA) or Jackson Laboratories (Bar Harbor, ME, USA). PLX-4720 was purchased from Med Chem Express (Monmouth Junction, NJ, USA) and resuspended in DMSO as the vehicle. OTX-008 was purchased from Axon Medchem (Reston, VA, USA). Purified Galectin-1 (cat: 10290-HNAE) was purchased from Sino Biological Europe GmbH (Eschborn, Germany).
3
Western Blot Analysis of Myogenic Markers
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The cells grown under each culture condition were washed by cold phosphate‐buffered saline and were directly harvested with 1× SDS sample buffer. The cells were sonicated by Branson Sonifier 450 (Branson Ultrasonics, Danbury, CT, USA). The homogenate (5–20 μL of each sample) was separated on SDS/PAGE gels for western blotting with anti‐DGKδ [9 (link)], anti‐cyclin D1 (sc‐450, Santa Cruz Biotechnology, Santa Cruz, CA, USA), anti‐cyclin D3 (sc‐182, Santa Cruz Biotechnology), anti‐myogenin (sc‐12732, Santa Cruz Biotechnology), anti‐myosin heavy chain (MyHC; MF20, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), anti‐β‐tubulin (T4026, Sigma–Aldrich), anti‐β‐actin (A5441, Sigma–Aldrich) and anti‐glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH; 016‐25523, Wako Pure Chemicals) antibodies. The immunoreactive bands were visualized using horseradish peroxidase‐conjugated anti‐rabbit or anti‐mouse IgG antibody (Cell Signaling Technology, Danvers, MA, USA) and Pierce™ ECL Western Blotting Substrate (Thermo Scientific, Waltham, MA, USA). The intensity of each band was measured using Amersham™ ImageQuant™ 800 (GE Healthcare, Chicago, IL, USA).
4
Western Blot Analysis of Tubulin and Aurora B
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Cells were washed with PBS, lysed with 1% SDS in PBS, and then incubated at 95 °C for 5 min. Sonication (Handy Sonic UR-20P) (TOMY SEIKO, Tokyo, Japan) was performed, and the lysate obtained was diluted using an SDS sample buffer (0.25 M Tris-HCl pH 6.8 containing 50% glycerol, 10% SDS, 0.025% bromophenol blue (BPB), 0.5 M dithiothreitol (DTT)), and the mixture was heated at 95 °C for 5 min. Proteins were separated by the SDS-PAGE and transferred onto a PVDF membrane, followed by blocking TBS-T containing 5% skim milk. Next, the membrane was incubated with a primary antibody solution diluted with 5% skim milk in TBS-T (in 0.02% NaH3) at 4 °C overnight. After washing with TBS-T, the membrane reacted with a secondary antibody solution diluted in TBS-T containing 5% skim milk for 30 min. After washing with TBS-T, the signal was detected using ECL Western Blotting Detection Reagents (GE Healthcare, Chicago, IL, USA) with the imaging analyzer LAS4000 (GE Healthcare, Chicago, IL, USA). The antibodies used in Western blotting were as follows: Anti-β-tubulin (T4026) (Sigma-Aldrich, St. Louis, MO, USA), anti-acetylated α-tubulin (T7451) (Sigma-Aldrich, St. Louis, MO, USA), anti-γ-tubulin (ab179503) (Abcam, Cambridge, UK), anti-Aurora B (611082) (BD, San Jose, CA, USA).
5
Metabolic Profiling using Extracellular Flux
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Oligomycin, carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP), rotenone, and antimycin A were used for metabolic measurement by extracellular flux analyzer (Agilent Technologies, CA, USA). Antibodies were as follows: anti-ZHX3 (ab9950) and anti-ACLY (ab40793) (Abcam, Cambridge, UK); anti-B23/NPM1 (sc-6013R, sc-271737), anti-mouse IgG (sc-2025) and anti-rabbit IgG (sc-2027) (Santa Cruz Biotechnology, Dallas, TX, USA); and anti-β-tubulin (T4026) (Sigma Aldrich, MO, USA).
6
Antibody Panel for Epigenetic Characterization
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The following antibodies were used in this study: anti‐Histone H3 tri‐methyl K27 (ab192985; Abcam, Cambridge, UK), anti‐EZH2 (clone AE25, cat. no. MABE362; Merck Millipore, Watford, UK) and anti‐β‐tubulin (T4026; Sigma‐Aldrich, Gillingham, UK) for immunoblotting; anti‐EZH2 for immunohistochemistry (anti‐EZH2, clone AE25, cat. no. MABE362, 1:1000; Merck Millipore, Watford, UK)38 ; anti‐γH2AX for immunofluorescence (05‐636‐AF555, 1:500; Merck Millipore, Watford, UK).
7
Cellular Protein Extraction and Western Blot
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Cells were plated in 6-well dishes. Lysates were prepared in lysis buffer (0.5% Triton X-100, 150 mM NaCl, 50 mM HEPES, 2 mM MgCl2, 2 mM EDTA, pH 7.4) via passage 10–15 times through a 26-gauge needle after sonication. After lysates were centrifuged at 13,000 ×g for 10 min at 4℃, the protein concentration in the supernatants was determined. The extracted proteins in sample buffer were loaded onto 10% Tris-glycine sodium dodecyl sulfate polyacrylamide gel electrophoresis gels. The proteins were transferred onto a polyvinylidene fluoride membrane. The membrane was incubated with anti-hemagglutinin (HA) (12CA5; Roche Molecular Biochemicals) and anti-β-tubulin (T-4026; Sigma Aldrich) overnight at 4℃.
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